LC-MS/MS method development and validation for the determination of polymyxins and vancomycin in rat plasma
LC-MS/MS method development and validation for the determination of polymyxins and vancomycin in rat plasma
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DOI：
10.1016/j.jchromb.2010.08.037
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发表时间：
2010-10-15
影响因子：
3
通讯作者：
Hansel, Steven
中科院分区：
文献类型：
--
作者：
Cheng, Chang;Liu, Shaorong;Hansel, Steven
建立了测定大鼠血浆中脂肽类多粘菌素和糖肽类万古霉素的液相色谱-串联质谱（LC-MS/MS）方法，并进行了验证。研究了三氯乙酸（TCA）浓度对样品回收率（从血浆中回收的样品峰面积/从纯溶剂溶液中回收的样品峰面积）的影响，确定了三氯乙酸（TCA）浓度为30%的最佳样品回收率。在Phenomenex Jupiter C18 5 μ 300埃50 mm × 2mm色谱柱上，采用pH为2.8的流动相，研究了TCA浓度对多肽色谱行为的影响。除了蛋白质沉淀。TCA也作为离子配对试剂，只存在于样品中，而不存在于流动相中。结果表明，随着TCA浓度的增加，分析物的保留率和灵敏度都有所提高。流动相中不含TCA有助于减少离子源污染，获得良好的重现性。血浆法对多粘菌素在5 ~ 5000 ng/mL范围内线性校准，精密度为2.3 ~ 10.8%，对多粘菌素B1、B2、E1、E2的精密度为91.7 ~ 107.4%。分别。对于万古霉素，校准范围为1至5000 ng/mL，精度为7.8-10.3，准确度为96.2-102.0%。多粘菌素B1、B2、E1、E2和万古霉素的loqs分别为7.5、18.1、7.3、5.0和1.0 ng/mL，变异系数小于20%。(C) 2010 Elsevier By。版权所有。
Simple, sensitive and robust liquid chromatography-tandem mass spectrometer (LC-MS/MS) methods were developed and validated for the determination of lipopeptide polymyxins and glycopeptide vancomycin in rat plasma. The effect of trichloroacetic acid (TCA) concentration on sample recoveries (peak area of sample recovered from plasma/peak area of sample from neat solvent solutions) was studied and an optimized concentration of 30% TCA were determined that gives the best sample recovery for the peptides from rat plasma. The effect of the TCA concentration on the chromatographic behavior of peptides was studied on a Phenomenex Jupiter C18 5 mu 300 angstrom 50 mm x 2 mm column using a mobile phase with a pH of 2.8. Other than protein precipitation. TCA also acted as ion pairing reagent and was only present in the samples but not in the mobile phases. The data demonstrated that by increasing the TCA concentration, the analyte retention and sensitivity were improved. The absence of TCA in mobile phase helped to reduce the ion source contamination and to achieve good reproducibility. The plasma method was linearly calibrated from 5 to 5000 ng/mL for polymyxins with precisions to be of 2.3-10.8%, and accuracies to be 91.7-107.4% for polymyxin B1, B2, E1, E2. respectively. For vancomycin the calibration is from 1 to 5000 ng/mL with precisions to be of 7.8-10.3 and accuracies to be 96.2-102.0%. The LLOQs corresponding with a coefficient of variation less than 20% were 7.5, 18.1, 7.3, 5.0 and 1.0 ng/mL for polymyxin B1, B2, E1, E2 and vancomycin, respectively. (C) 2010 Elsevier By. All rights reserved.